Covalent Portal Protein Tethering for Stable Solid-State Nanopores
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Solution Overview
Problem
Existing nanopore technologies face challenges in achieving reproducible, stable, and uniformly oriented protein immobilization within solid-state nanopores, limiting their voltage range and durability, especially when combining protein and solid-state pores in hybrid systems.
Innovation Solution
A method involving the covalent immobilization of the G20c portal protein within a chemically modified SiNx nanopore through disulfide bonding between cysteine residues of the protein and a thiol-silane surface, using 2,2-dimethoxyl-1-thia-2-silacyclopentane, allows for stable hybrid nanopores functional at high voltages.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If protein nanopores are used, then recognition and quantification performance is improved, but chemical and physical robustness deteriorates due to organic membrane instability
Solution Approach 1:
The patent merges protein nanopores with solid-state nanopores to create a hybrid system that combines the measurement precision advantages of protein pores with the robustness of solid-state membranes, resolving the contradiction between performance and reliability
Solution Approach 2:
The hybrid nanopore system uses composite material structure combining organic protein components with inorganic solid-state membrane, achieving both high measurement precision and chemical-physical robustness simultaneously
2Reliability
If solid-state nanopores are used, then chemical and physical robustness is improved, but ability to reproduce atomic detail deteriorates
Solution Approach 1:
The patent segments the nanopore system into two functional parts: solid-state membrane providing robustness and protein nanopore providing atomic-level structural precision, allowing each component to excel at its specialized function
3Ease of manufacture
If hybrid pores are formed without covalent linking, then ease of manufacture is improved, but stability and voltage range limitation worsens
Solution Approach 1:
The patent introduces covalent bonds as an intermediary mechanism to chemically link the protein nanopore to the solid-state membrane, providing stable attachment that enables high-voltage operation while maintaining relatively simple fabrication procedures
4Measurement precision
If non-specific binding is eliminated, then measurement precision is improved, but understanding of membrane-protein interaction complexity increases
Solution Approach 1:
The patent applies local quality modification by functionalizing specific regions of the solid-state membrane with chemical groups that selectively interact with the protein nanopore, eliminating non-specific binding while maintaining controlled, predictable interactions
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The hybrid nanopore system enables high-voltage sensing and motor protein-mediated ratcheting, facilitating long-range DNA scanning and improved pore resolution, overcoming limitations of traditional membrane-based pores.
Implementation Method 1
forming a disulfide bond between a cysteine residue of the portal protein and the thiol-silane surface of the membrane comprising the nanopore
Data Source
AI summary
Sensors and related methods of making and using sensors are disclosed. The sensor include a portal protein, such as G20c derived from the bacteriophage Thermits thermophilus, covalently bound to a thiol-silane surface of a solid-state membrane having one or more nanopores. The sensors can be used for protein and/or nucleotide sequencing and for small-molecule detection.